Purpose :
Differentiation of corneal fibroblasts to myofibroblasts is a major underlying mechanism for corneal scarring development. Inhibitor of differentiation (Id) genes regulate cellular proliferation, differentiation and fibrosis in many tissues. Recently, we characterized Id genes expression in human cornea, and their role in corneal wound healing. This study tested the hypothesis that Id2 or Id3 over-expression into human corneal fibroblasts effectively blocks differentiation of corneal fibroblasts to myofibroblasts and offers an innovative gene therapy method to treat corneal fibrosis in vivo.

Methods :
Primary human corneal fibroblast (HCF) and corneal myofibroblast (HMF) cultures generated from donor human corneas were used. HMF were produced by growing HCF in 5ng/ml TGFβ1 for 72h under serum-free conditions. Lipofectamine-3000 and G418 were used for gene transfer and stable clone selection, respectively. Immunofluorescence, immunoblotting, and qPCR were used to confirm gene transfer and quantify mRNA and protein levels of profibrotic markers in cultures. New Zealand White rabbits were used for in vivo studies.